Journal article
Enzyme-linked immunosorbent assay for detection of organophosphorylated butyrylcholinesterase: A biomarker of exposure to organophosphate agents
Analytica chimica acta, Vol.693(1), pp.1-6
2011
Handle:
https://hdl.handle.net/2376/106990
PMID: 21504805
Abstract
A sandwich enzyme-linked immunosorbent assay (sELISA) has been developed for detection of organophosphorylated butyrylcholinesterase (OP-BChE), a potential biomarker for human exposure to organophosphate insecticides and nerve agents. A pair of antibodies specific to OP-BChE adduct were identified through systematic screening of several anti BChE antibodies (anti-BChE) and anti-phosphoserine antibodies (anti-P
ser) from different sources. The selected anti-BChE (set as capture antibody) antibodies recognize both phosphorylated and nonphosphorylated BChE. These antibodies can therefore be used to capture both BChE and OP-BChE from the sample matrices. The anti-P
ser (set as detecting antibody) was used to recognize the OP moiety of OP-BChE adducts. With the combination of the selected antibody pair, several key parameters (such as the concentration of anti-BChE and anti-P
ser, and the blocking agent) were optimized to enhance the sensitivity and selectivity of the sELISA. Under the optimal conditions, the sELISA has shown a wide linear range from 0.03
nM to 30
nM, with a detection limit of 0.03
nM. Furthermore, the sELISA was successfully applied to detect OP-BChE using
in vitro biological samples such as rat plasma spiked with OP-BChE with excellent adduct recovery (
z
>
99%). These results demonstrate that this novel approach holds great promise to develop an ELISA kit and offers a simple and cost-effective tool for screening/evaluating exposure to organophosphate insecticides and nerve agents.
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Details
- Title
- Enzyme-linked immunosorbent assay for detection of organophosphorylated butyrylcholinesterase: A biomarker of exposure to organophosphate agents
- Creators
- Limin Wang - Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDan Du - Pacific Northwest National Laboratory, Richland, WA 99352, United StatesDonglai Lu - Pacific Northwest National Laboratory, Richland, WA 99352, United StatesChiann-Tso Lin - Pacific Northwest National Laboratory, Richland, WA 99352, United StatesJordan N Smith - Pacific Northwest National Laboratory, Richland, WA 99352, United StatesCharles Timchalk - Pacific Northwest National Laboratory, Richland, WA 99352, United StatesFengquan Liu - Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaJun Wang - Pacific Northwest National Laboratory, Richland, WA 99352, United StatesYuehe Lin - Pacific Northwest National Laboratory, Richland, WA 99352, United States
- Publication Details
- Analytica chimica acta, Vol.693(1), pp.1-6
- Academic Unit
- School of Mechanical and Materials Engineering
- Publisher
- Elsevier B.V
- Identifiers
- 99900546712701842
- Language
- English
- Resource Type
- Journal article